Characteristics of earthworms (Eisenia fetida) in PAHs contaminated soil amended with sewage sludge or vermicompost

Earthworms can be used to remove polycyclic aromatic hydrocarbons (PAHs) from soil, but this might affect their survival and they might accumulate the contaminants. Sterilized and unsterilized soil was contaminated with phenanthrene (Phen), anthracene (Anth) and benzo(a)pyrene (BaP), added with or without Eisenia fetida, sewage sludge or vermicompost. Survival, growth, cocoon formation and concentrations of PAHs in the earthworms were monitored for 70 days. Addition of sewage sludge to sterilized or unsterilized soil maintained the number of earthworms and their survival was 94%. The addition of sludge significantly increased the weight of earthworms 1.3 times compared to those kept in the unamended soil or in soil amended with vermicompost. The weight of earthworms was significantly lower in sterilized than in unsterilized soil. Cocoons were only detected when sewage sludge was added to unsterilized soil. A maximum concentration of 62.3 μg Phen kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 7 days and 22.3 μg Phen kg⁻¹ when kept in the unamended unsterilized soil after 14 days. Concentrations of Phen in the earthworms decreased thereafter and ≤2 μg kg⁻¹ after 28 days. A maximum Anth concentration of 82.5 μg kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost and 45.8 μg Anth kg⁻¹ when kept in the unamended unsterilized soil after 14 days. A maximum concentration of 316 μg BaP kg⁻¹ was found in the earthworms kept in sterilized soil amended with vermicompost after 56 days and 311 μg BaP kg⁻¹ when kept in the unsterilized soil amended with vermicompost after 28 days. The amount of BaP in the earthworm was generally largest after 28 days, but after 70 days still 60 μg kg⁻¹ was found in E. fetida when kept in the sterilized soil amended with sewage sludge. It was found that E. fetida survived in PAHs contaminated soil and accumulated only small amounts of the contaminants, but sewage sludge was required as food for its survival and cocoon production.     

Inhibition of the germination and growth of Phymatotrichopsis omnivora (cotton root rot) by oilseed meals and isothiocyanates

The meals (co-products remaining after oil extraction) from many oilseed crops contain biocidal chemicals that are known to inhibit the growth and activity of some soil microorganisms including several plant pathogens. The fungus Phymatotrichopsis omnivora (Duggar) Hennebert is the causal agent of cotton root rot that has greatly hindered the production of cotton and alfalfa in Texas and the southwestern USA. We investigated the effect of oilseed meals from both brassicaceous plants including mustard and camelina as well as non-brassicaceous plants including jatropha, flax, and Chinese tallow on P. omnivora sclerotial germination and hyphal growth in Branyon clay soil, as well as the effects of 4 types of individual isothiocyanates (ITCs) including allyl, butyl, phenyl, and benzyl ITC on P. omnivora growth on potato dextrose agar (PDA). The oilseed meals were added to the soil at rates of 0%, 1%, and 5% (w/w). The results showed that all tested brassicaceous and jatropha seed meals were able to inhibit P. omnivora sclerotial germination and hyphal growth at 5% and 1% application rates respectively, with mustard seed meal being the most effective. Neither flax nor Chinese tallow showed any inhibitory effects on sclerotial germination. All tested ITCs inhibited P. omnivora OKAlf8 hyphal growth, although the level of inhibition varied with concentration. The IC₅₀ values were 0.62 ± 0.19, 4.47 ± 0.08, 5.67 ± 0.10, and 20.48 ± 0.30 μg cm⁻³ for allyl, butyl, phenyl, and benzyl ITC respectively. These results indicate that press meals of several brassicaceous species as well as jatropha may have potential for reducing cotton root rot.     

Microbial utilization and mineralization of [14C]glucose added in six orders of concentration to soil

The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO₂ efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 μM in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding ¹⁴C-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO₂ was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 μg glucose C g^?1 soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 μg glucose C g^?1 soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 μg C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between micro-organisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 μg C g^?1). At glucose additions higher than 2.6 μg C g^?1 soil, however, the increase of the glucose mineralization per 1 μg added glucose was much smaller as at additions below 2.6 μg C g^?1 soil and was accompanied by decreasing portions of glucose ¹⁴C incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6–257 μg C g^?1 (=0.78–78% of microbial biomass C). At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO₂. The addition of 257 μg glucose C g^?1 soil (0.78 μg C glucose μg^?1 C micro-organisms) caused a lag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics.     

Plant growth promoting potential of Pontibacter niistensis in cowpea (Vigna unguiculata (L.) Walp.)

During the past couple of decades, understanding of rhizosphere biology has progressed with the discovery of a special group of microorganisms known as plant growth promoting rhizobacteria (PGPR) and its application for sustainable agriculture has increased tremendously in various parts of the world. The search for microorganisms that improve soil fertility and enhance plant nutrition has continued to attract attention due to the increasing cost of fertilizers and some of their negative environmental impacts. In this study we demonstrated, a novel bacterial species Pontibacter niistensis NII-0905 isolated from forest soil in Western ghat forest soil with potential plant growth promoting ability (PGP) such as phosphate solubilization, indole acetic acid (IAA), siderophore and hydrogen cyanide (HCN) production. The activity varies with different growth temperatures, strain solubilize 28.5 ± 0.9, 48.02 ± 1.9 and 65.07 ± 2.1 μg mL⁻¹ at 4, 15 and 30 °C respectively and produced 24.8 μg mL⁻¹ day⁻¹ of indole acetic acid (IAA) in tryptophan amended media. Qualitative detection of siderophore production and HCN were also detected at all temperature tested. At a lower temperature (4 °C) strain NII-0905 retained all the plant growth promotion attributes. A significant increase in the growth of cow pea was recorded with inoculations of strain NII-0905 in pot experiments. Scanning electron microscopic study revealed the root colonization on cow pea seedlings against the untreated one. These results demonstrate that, the isolate NII-0905 has the promising PGPR attributes for both in cold as well as in humid condition. It has potential as a biofertilizer to enhance soil fertility and promote the plant growth.     

Formation and use of microbial residues after adding sugarcane sucrose to a heated soil devoid of soil organic matter

A 67-day incubation experiment was carried out with a soil initially devoid of any organic matter due to heating, which was amended with sugarcane sucrose (C₄-sucrose with a δ¹³C value of ?10.5‰), inorganic N and an inoculum for recolonisation and subsequently at day 33 with C₃-cellulose (δ¹³C value of ?23.4‰). In this soil, all organic matter is in the microbial biomass or in freshly formed residues, which makes it possible to analyse more clearly the role of microbial residues for decomposition of N-poor substrates. The average δ¹³C value over the whole incubation period was ?10.7‰ in soil total C in the treatments without C₃-cellulose addition. In the CO₂ evolved, the δ¹³C values decreased from ?13.4‰ to ?15.4‰ during incubation. In the microbial biomass, the δ¹³C values increased from ?11.5‰ to ?10.1‰ at days 33 and 38. At day 67, 36% of the C₄-sucrose was left in the treatment without a second amendment. The addition of C₃-cellulose resulted in a further 7% decrease, but 4% of the C₃-cellulose was lost during the second incubation period. Total microbial biomass C declined from 200 μg g^?1 soil at day 5 to 70 μg g^?1 soil at day 67. Fungal ergosterol increased to 1.5 μg g^?1 soil at day 12 and declined more or less linearly to 0.4 μg g^?1 soil at day 67. Bacterial muramic acid declined from a maximum of 35 μg g^?1 soil at day 5 to a constant level of around 16 μg g^?1 soil. Glucosamine showed a peak value at day 12. Galactosamine remained constant throughout the incubation. The fungal C/bacterial C ratio increased more or less linearly from 0.38 at day 5 to 1.1 at day 67 indicating a shift in the microbial community from bacteria to fungi during the incubation. The addition of C₃-cellulose led to a small increase in C₃-derived microbial biomass C, but to a strong increase in C₄-derived microbial biomass C. At days 45 and 67, the addition of N-free C₃-cellulose significantly decreased the C/N ratio of the microbial residues, suggesting that this fraction did not serve as an N-source, but as an energy source.     

Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

Food choice and reproductive success of Folsomia candida feeding on copper-contaminated mycelium of the soil fungus Alternaria alternata

We examined collembolan food preference for fungal mycelium grown on copper-contaminated medium, and the relationship between copper content, food selectivity and collembolan fitness when fed contaminated mycelium.To clarify whether collembolan food selectivity is related to fitness parameters, Folsomia candida were fed mycelium of the dark-pigmented fungus Alternaria alternata grown on medium with different copper concentrations. Copper-contaminated food (fungus grown on 50, 125, 250 and 500 μg Cu g^?1 medium, fresh wt.) was offered together with untreated food for 4 weeks. F. candida fed selectively on the provided mycelium and discriminated clearly between mycelium grown on high and low levels of contamination, distinctly preferring fungus grown on medium with a total copper concentration of 50 and 125 μg g^?1. In contrast, fungus grown on highly contaminated medium (250 and 500 μg g^?1) was avoided. Collembolan food preference generally matched fitness parameters. Reproduction was significantly affected by the total copper concentration of the fungal growth medium. When fed their preferred mycelium, collembolan reproduction was enhanced, whereas a diet of highly contaminated mycelium (250 or 500 μg g^?1) resulted in a strong decrease in reproduction. Adult survival was affected only marginally. Even though heavy metal contamination is a potential stress factor for many soil microarthropods, F. candida is able to discriminate between high and low quality food sources, and even benefits from moderately elevated copper concentrations.     

Tolerance,growth and degradation of phenanthrene and benzo[a]pyrene by Rhizobium tropici CIAT 899 in liquid culture medium

Polycyclic aromatic hydrocarbons (PAH) are ubiquitous pollutants that are toxic and recalcitrant to degradation by bacteria. This research evaluated the toxicity of different concentrations [10, 20, 40, 60, 80 and 100 μg mL⁻¹] of phenanthrene (PHE) or benzo[a]pyrene (BaP) on the growth of Rhizobium tropici CIAT899 under in vitro conditions as well as the potential degradation of PHE and BaP by this bacterium. At 24 h, a 40% decrease in Rhizobium growth was observed when exposed to 40 μg mL⁻¹ of either PHE or BaP. Furthermore, bacterial growth was completely inhibited by PHE or BaP applied in 80 and 100 μg mL⁻¹. After 96 h, the growth of R. tropici at 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ was similar to those treatments without PAH. To evaluate R. tropici degrading capabilities, supernatants of cultures with 40 μg PHE mL⁻¹ or 60 μg BaP mL⁻¹ were analyzed by gas chromatography coupled to mass spectrophotometer (GC–MS). R. tropici was able to degrade either PHE or BaP diminishing its concentration in 20% and 25% during the first 24 h, degradation obtained at 120 h was 50% and 45% for PHE or BaP, respectively. This research shows for the first time that R. tropici CIAT 899 grows in liquid culture medium contaminated with PAH, and moreover is able to growth and to degrade either PHE or BaP.     

Improvement of Brassica napus growth under cadmium stress by cadmium-resistant rhizobacteria

This study focuses on the characterization of four bacterial isolates from heavy metal-polluted rhizosphere in order to examine their plant growth promoting (PGP) activity. The PGP activity on the canola (Brassica napus) of the strains which showed cadmium resistance and multiple PGP traits was assessed in the presence and in the absence of Cd²⁺. The strains, Pseudomonas tolaasii ACC23, Pseudomonas fluorescens ACC9, Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14 showed 1-aminocyclopropane-1-carboxylate deaminase (ACCD) activity. They also synthesized ACCD enzyme in vitro when 0.4 mM Cd²⁺ was added to the growth medium. The presence of the metal, however, reduced the ACCD activity in Alcaligenes sp. ZN4 and Mycobacterium sp. ACC14, while it did not affect the ACCD activity of P. tolaasii ACC23 and P. fluorescens ACC9. ACC9 and ACC23 produced indole acetic acid (IAA) and siderophores, while ACC14 produced only IAA. IAA and siderophores were produced more actively under Cd-stress.Root elongation assays conducted on B. napus under gnotobiotic conditions demonstrated increases (from 34% up to 97%) in root elongation of inoculated canola seedlings compared to the control plants. Subsequently, the effect of inoculation with these strains on growth and uptake of Cd²⁺ in roots and shoots of canola was studied in pot experiments using Cd-free and Cd-treated (15 μg Cd²⁺ g^?1 dw) soil. Inoculation with P. tolaasii ACC23, P. fluorescens ACC9 and Mycobacterium sp. ACC14 promoted the growth of plants at concentrations of 0 and 15 μg Cd²⁺ g^?1 soil. The maximum growth was observed in the plants inoculated with P. tolaasii ACC23. The strains did not influence the specific accumulation of cadmium in the root and shoot systems, but all increased the plant biomass and consequently the total cadmium accumulation.The present observations showed that the bacterial strains used in this study protect the plants against the inhibitory effects of cadmium, probably due to the production of IAA, siderophores and ACCD activity.     

Botanical survey and screening of plant species which accumulate 226Ra from contaminated soil of uranium waste depot

A geobotanical study was performed of the wild plants growing in the area of the old uranium mill tailings waste depot of a former uranium ore reprocessing factory in South Bohemia and the distribution of ²²⁶Ra in selected plants was determined. The distribution of ²²⁶Ra in contaminated soil was found to be extremely variable (from 7 to 32 Bq ²²⁶Ra g⁻¹ of DW). The differences in plant distribution were caused by factors of disturbance, soil properties (nutrients and salt content, water supply), and successional stage. No direct relation was proved between plant species characteristics and their radioactivity content. The results showed a great range of variation in the accumulation of ²²⁶Ra by the plant species found. The highest activity of ²²⁶Ra was found in Potentilla reptans (4.09 Bq ²²⁶Ra g⁻¹ of DW), Mentha arvensis (4.00 Bq ²²⁶Ra g⁻¹ of DW), and Daucus carota (3.70 Bq ²²⁶Ra g⁻¹ of DW). About half of the plant species are used as medicinal plants and some of them are accumulators of ²²⁶Ra. However, no plants suitable for phytoextraction of ²²⁶Ra contaminated substrates were discovered.     

Enzyme activities and diuron persistence in soil amended with vermicompost derived from spent grape marc and treated with urea

Mineral fertilizers, organic amendments, and pesticides are inputs commonly used in conventional farming practices. The aim of this study was to evaluate the effects of single or combined applications of spent grape marc-vermicompost, urea, and/or diuron on soil-enzyme activities and the persistence of this herbicide in soils with low organic carbon content. The application of vermicompost enhanced dehydrogenase (DHase) enzyme activity over time but altered soil urease activity to a very limited extent. The reduction in diuron concentrations and the increase in DHase activity indicated that the soil microorganisms were capable of degrading the ureic herbicide. Treatment with vermicompost and diuron had a stimulatory effect on soil microbial activity. On the whole, the application of diuron and urea to the vermicompost-amended soil raised DHase and urease activity to maximum levels (>3 μg INTF g^?1 h^?1 and >47 μg NH₄⁺ g^?1 h^?1, respectively). The application of urea to the unamended and vermicompost-amended soil decreased diuron persistence from 18.8 and 33 d to 12.5 and 15 d, respectively. Our findings show that although vermicompost additions reduce diuron availability, this boosts diuron degradation when combined with urea. These additions, under different soil management conditions, minimize the bioavailability and persistence of diuron and consequently the risk of leaching and seepage into aquifers. Compared with untreated soils, these types of treated soils could also improve agricultural sustainability and the quality of the environment.     

Impact of carbon-rich dairy factory effluent on growth of perennial ryegrass (Lolium perenne) and soil microorganisms

A pot experiment was conducted to investigate the impact of high carbon dairy factory effluent application on the growth of perennial ryegrass (Lolium perenne L.), plant nutrient uptake, soil microbial biomass carbon and nitrogen, populations of soil-microorganisms, root colonising fungi and the microbial functional diversity. The effluent was added at rates of 0, 100,000, 200,000 and 300,000 l ha^–1. These rates are equivalent to 0, × 1, × 2 and × 3 normal field application rates. The added effluent contained (g l^–1), C; 19.42, total P; 0.65; S, 0.75, K; 1.33, Na; 4.55, Mg; 0.11, NH₄; 0.073, total N; 0.073 and had a pH of 4.33. Replicate pots (incubated in a controlled-environment room at 20 °C, with 16 h light/8 h dark) were harvested at 32, 61, and 130 days after setting up of the experiment. In the first sampling, shoot dry matter levels declined significantly (P < 0.01) with increased effluent. By the third sampling the trend was reversed with treated pots having greater amounts of shoot dry matter. The initial depression of growth was possibly due to a combination of factors including excess levels of available carbon (C) for microbes leading to immobilisation of nutrients, particularly nitrogen (N) and sulphur (S). Shoot N and S concentrations were lower (P < 0.001) and the phosphorus concentrations were higher in effluent-treated samples. Soil microbial biomass-C (480 and 770 μg g⁻¹ of biomass C in untreated and treated soil, respectively) and microbial-N (81 and 123 μg g^–1 of microbial-N in untreated and treated soil, respectively) were significantly (P < 0.001) greater in effluent-treated pots at all times. Populations of total culturable bacteria were higher (P < 0.01) in the treated pots in the first sample (log₁₀ populations g^–1 were 7.3 in untreated pots compared to 8.0 averaged across three treatments) but there were no differences in the subsequent two samples. Effluent also increased yeast populations (log₁₀ numbers g^–1 were 0.6 in untreated pots and 3.1 in treated pots averaged across treatments and times P < 0.01) at all three sampling times. The Shannon-Weiner Diversity Index of root fungi decreased with increasing effluent application (P < 0.01) while the species richness decreased with effluent as well as with time (P < 0.1). Potential root pathogens Fusarium oxysporum, total Fusarium spp. and Pythium spp. significantly increased (P < 0.05) in treated samples but in the final sampling, Codinaea fertilis significantly (P < 0.05) decreased with effluent treatment. The microbial functional diversity pattern and the average well colour development (AWCD) in soil were significantly changed by the effluent application but effects were not detectable after 130 days.     

Emission of nitrous oxide from hydrocarbon contaminated soil amended with waste water sludge and earthworms

Soils in Mexico are often contaminated with hydrocarbons and addition of waste water sludge and earthworms accelerates their removal. However, little is known how contamination and subsequent bioremediation affects emissions of N₂O and CO₂. A laboratory study was done to investigate the effect of waste water sludge and the earthworm Eisenia fetida on emission of N₂O and CO₂ in a sandy loam soil contaminated with the polycyclic aromatic hydrocarbons (PAHs): phenanthrene, anthracene and benzo(a)pyrene. Emissions of N₂O and CO₂, and concentrations of inorganic N (ammonium (NH₄⁺), nitrite (NO₂^?) nitrate (NO₃^?)) were monitored after 0, 5, 24, 72 and 168 h. Adding E. fetida to the PAHs contaminated soil increased CO₂ production rate significantly 2.0 times independent of the addition of sludge. The N₂O emission rate from unamended soil expressed on a daily base was 5 μg N kg^?1 d^?1 for the first 2 h and increased to a maximum of 325 μg N kg^?1 d^?1 after 48 h and then decreased to 10 μg N kg^?1 d^?1 after 168 h. Addition of PAHs, E. fetida or PAHs + E. fetida had no significant effect on the N₂O emission rate. Adding sludge to the soil sharply increased the N₂O emission rate to >400 μg N kg^?1 d^?1 for the entire incubation with a maximum of 1134 μg N kg^?1 d^?1 after 48 h. Addition of E. fetida, PAHs or PAHs + E. fetida to the sludge-amended soil reduced the N₂O emission rate significantly compared to soil amended with sludge after 24 h. It was found that contaminating soil with PAHs and adding earthworms had no effect on emissions of N₂O. Emission of N₂O, however, increased in sludge-amended soil, but addition of earthworms to this soil and contamination reduced it.     

The fate and toxicity of the flavonoids naringenin and formononetin in soil

The flavonoid class of plant secondary metabolites play a multifunctional role in below-ground plant–microbe interactions with their best known function as signals in the nitrogen fixing legume–rhizobia symbiosis. Flavonoids enter rhizosphere soil as a result of root exudation and senescence but little is known about their subsequent fate or impacts on microbial activity. Therefore, the present study examined the sorptive behaviour, biodegradation and impact on dehydrogenase activity (as determined by iodonitrotetrazolium chloride reduction) of the flavonoids naringenin and formononetin in soil. Organic carbon normalised partition coefficients, log K_oc, of 3.12 (formononetin) and 3.19 (naringenin) were estimated from sorption isotherms and, after comparison with literature log K_oc values for compounds whose soil behaviour is better characterised, the test flavonoids were deemed to be moderately sorbed. Naringenin (spiked at 50 μg g^?1) was biodegraded without a detectable lag phase with concentrations reduced to 0.13±0.01 μg g^?1 at the end of the 96 h time course. Biodegradation of formononetin proceeded after a lag phase of ～24 h with concentrations reduced to 4.5±1% of the sterile control after 72 h. Most probable number (MPN) analysis revealed that prior to the addition of flavonoids, the soil contained 5.4×10⁶ MPN g^?1 (naringenin) and 7.9×10⁵ MPN g^?1 (formononetin) catabolic microbes. Formononetin concentration had no significant (p>0.05) effect on soil dehydrogenase activity, whereas naringenin concentration had an overall but non-systematic impact (p=0.045). These results are discussed with reference to likely total and bioavailable concentrations of flavonoids experienced by microbes in the rhizosphere.     

Miscanthus sinensis and wild flowers as food resources of Lumbricus terrestris L.

Senescent leaves of Miscanthus sinensis contained 36% soluble polysaccharides, 26% cellulose and had a C/N ratio of 45. In 11 wild flower species contents of soluble polysaccharides (21–30%), cellulose (3–16%) and C/N ratio (13–31) were lower. Decomposing leaves of M. sinensis lost weight at a rate of 0.002 day⁻¹, increased the C/N ratio from 45 to about 100, the bacterial biomass from 0.4 to 1 μg C mg⁻¹ dry weight, and decreased the tensile strength from 35 to 10 N. The withdrawal rate of Lumbricus terrestris with senescent leaves of M. sinensis was 30 mg g⁻¹ week⁻¹; the feeding rate was lower. With most senescent wild flowers withdrawal and feeding rates were higher. During decomposition of M. sinensis withdrawal rates increased to about 90, and feeding rates to about 30 mg g⁻¹ week⁻¹. The rates were not related to soluble polysaccharides, cellulose, acid-insoluble residue, C/N ratio and the presence of trichomes on the leaves. The abundance of L. terrestris decreased in a meadow turned into a field of M. sinensis from 55 to 26 earthworms m⁻² and increased in a rotational maize field turned into wild flower strips from 28 to 46 earthworms m⁻². The species richness of earthworms decreased with M. sinensis from 7.2 to 4.7 and increased with wild flowers from 4.7 to 6.7 species per sampling unit.     

Aggregate water-stability,particle-size and soil solution properties in conducive and suppressive soils to Fusarium wilt of banana from Canary Islands (Spain)

The influence of several soil properties on soil conduciveness or suppressiveness to disease caused by the soil fungus Fusarium oxysporum f. sp. cubense was studied in seven field plots of banana plantations, situated in Tenerife and Gran Canaria islands (Canary Islands, Spain). In each plot, soil samples were taken in conducive and suppressive areas to Fusarium wilt. Water-stable aggregates (WSA: 200–2000 μm diameter), soil particle size, and selected soil solution characteristics [pH, electric conductivity (EC) and soluble Na] were determined in the samples. Aggregate water-stability was higher in soils of conducive areas than in suppressive areas. The percentage of WSA in the conducive areas ranged from 460 to 330 g kg⁻¹, while in the suppressive areas the maximum value was 285 g kg⁻¹ and the minimum was 150 g kg⁻¹. The soils had high clay content and the EC and soluble Na tended to be higher in suppressive areas than in conducive areas. Soil solution pH was lower in conducive areas (except sites 1 and 9). Our data provide evidence that in different soil areas of the same plot, the structural stability of aggregates, presumably controlled in part by the clay fraction, soluble Na concentration and EC, is of great importance for the conduciveness or suppressiveness to banana wilt caused by Fusarium oxysporum f. sp. cubense of the soils studied. Finally, we hypothesize that a greater stability of the aggregates forming anaerobiosis could partly explain most of the available Fe found in soil areas where the disease was severe, at least in these types of soils.     

Estimation of chlorophyll content in Eucalyptus globulus foliage with the leaf reflectance model PROSPECT

This communication presents the performance of the PROSPECT leaf optical model to derive chlorophyll content (Cab) estimates from reflectance and transmittance spectra of Eucalyptus globulus foliage. Estimates were compared to measured chlorophyll of 100% acetone extractions. The analysis showed that recent modifications to the absorption coefficients used in the PROSPECT model resulted in improved estimates of chlorophyll for both adult and juvenile leaves. Results were better for adult leaves, with estimates within 5 μg cm^?2 for the juvenile data set and 3 μg cm^?2 for adult leaves. Accurate estimates of canopy chlorophyll content in eucalypt plantations through the numerical inversion of leaf-level radiative transfer (RT) models such as PROSPECT coupled to canopy RT models would improve our ability to assess and monitor eucalypt plantation growth, health and condition.     

Short-term CO2 emissions from planted soil subject to elevated CO2 and simulated precipitation

Carbon dioxide emissions from soils beneath canopies of two Mediterranean plants, Artemisia absinthium L. and Festuca pratensis Huds. cv. Demeter, were monitored over a 7-day period that included an artificial precipitation event of 4 cm. The experiments were conducted using 0.2 m³ soil microcosms inside greenhouses with CO₂ concentrations of either 360 or 500 μmol mol⁻¹. Carbon dioxide flux from the soil surface, as calculated using a diffusive transport model agreed well with CO₂ flux measurements made using a dynamic flow system. Soil CO₂ emissions did not differ significantly between the 360 and 500 μmol mol⁻¹ CO₂ treatments when soils were dry (volumetric soil moisture content ≤9%). A simulated precipitation event caused an immediate exhalation of CO₂ from soil, after which CO₂ emissions declined slightly and remained constant for approximately 36 h. CO₂ emissions from soil microcosms with F. pratensis plants growing in 500 μmol mol⁻¹ CO₂ then rose to levels that were significantly greater than CO₂ emissions from soils in the microcosms exposed to 360 μmol mol⁻¹ CO₂. For A. absinthium growing in 500 μmol mol⁻¹ CO₂, the rise in soil CO₂ emissions following the wetting event was not significantly greater than emissions from soils with A. absinthium growing under 360 μmol mol⁻¹ CO₂. A. absinthium above ground biomass increased by 46.1 ± 17.9% (mean ± S.E., n = 4, P ≤ 0.05). Above ground biomass did not significantly increase for F. pratensis (14.4 ± 6.5%, P ≥ 0.10). Root biomass, on the other hand, increased for both species; by 50.6 ± 17.9% (P ≤ 0.05) for A. absinthium and by 55.9 ± 12.7% (P ≤ 0.05) for F. pratensis. Our results demonstrate two events following precipitation onto dry soils, an immediate release of CO₂ followed by a gradual increase from enhanced biological activity The gradual increase was greater for the herbaceous ruderal perennial F. pratensis under elevated CO₂.     

The role of biological activity (roots,earthworms) in medium-term C dynamics in vertisol under a Digitaria decumbens (Gramineae) pasture

The natural abundance of ¹³C was used to estimate the turnover of the soil organic matter in a vertisol re-grassed with Digitaria decumbens (C4 plant) following intensive market gardening (C3 plants). In addition, the experimental design allowed us to determine the respective roles of roots and earthworms (Polypheretima elongata) in soil C stock restoration in D. decumbens pasture.The C stock increased from 31 to 37 Mg C ha⁻¹ in 5 years and the δ¹³C increased from −18.1‰ in market gardening soil to −15.5‰ in the 5-year-old pasture soil in the upper 20 cm. Below the 20 cm soil layer, the C stock and the δ¹³C did not change significantly in 5 years. The net gain of 6 Mg C ha⁻¹ was the balance of a loss of 5 Mg C ha⁻¹ derived from market gardening and a gain of 11 Mg C ha⁻¹ derived from D. decumbens. Effects of earthworms on the C dynamics were not discernible.     

Field margins and management affect settlement and spread of an introduced dew-worm (Lumbricus terrestris L.) population

To study the feasibility of earthworm introduction for increasing the macroporosity and permeability of arable heavy clay, deep-burrowing earthworms (Lumbricus terrestris L.) were inoculated into a tile drained experimental field in Jokioinen, S-W Finland in autumn 1996. Inoculation with the Earthworm Inoculation Unit technique was at the up-slope end of the field, in the field margins under permanent grass, and inside the four 0.46 ha plots of the field. The experiment was monitored on three occasions. In 1998 the L. terrestris population had persisted in low numbers only in field and plot margins. By 2003, when the field had been under set-aside grass for three years, density had grown in the margins and L. terrestris were also found inside the field at a very low density. The third monitoring was in autumn 2009, after a further four years as set-aside and a subsequent division of the field into no-till and ploughing management, and looked at the effects of management (margins, no-till, ploughing), distance from the inoculation and sub-drainage on L. terrestris abundance. The abundance displayed a clear gradient over the field, declining from 14 ind. and 18 g m^?2 at 5–9 m from inoculation, to 1 ind. and 2 g m^?2 at 56–60 m distance. Margins had the highest abundances (16 ind. and 32 g m^?2), followed by no-till (4 ind. and 4 g m^?2) and ploughing (1 ind. and 1 g m^?2). Abundances were significantly higher above the tiles than between them (P < 0.05). The results demonstrate the importance of no-till and sub-drain line habitats as settlement supports for the inoculated population. Field margins proved to be decisive for inoculation success, by providing bridgeheads for population establishment and later by acting as source areas for the colonisation of the field. This finding highlights the general importance of field margins in the dispersal ecology of earthworms in arable landscapes.